流感嗜血杆菌中必需基因的鉴定与分析。

Identification and analysis of essential genes in Haemophilus influenzae.

作者信息

Wong Sandy M S, Akerley Brian J

机构信息

Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, MA, USA.

出版信息

Methods Mol Biol. 2008;416:27-44. doi: 10.1007/978-1-59745-321-9_3.

DOI:10.1007/978-1-59745-321-9_3

PMID:18392959

Abstract

The human respiratory pathogen Haemophilus influenzae, a Gram-negative bacterium, is the first free-living organism to have its complete genome sequenced, providing the opportunity to apply genomic-scale approaches to study gene function. This chapter provides an overview of a highly efficient, in vitro mariner transposon-based method that exploits the natural transformation feature of this organism for the identification of essential genes. In addition, we describe strategies for conditional expression systems that would facilitate further analysis of this class of genes. Finally, we outline a method based on the approach used in H. influenzae for identifying essential genes that can be applied to other bacteria that are not naturally transformable.

摘要

人类呼吸道病原体流感嗜血杆菌是一种革兰氏阴性菌，它是首个完成全基因组测序的自由生活生物体，这为应用基因组规模的方法研究基因功能提供了契机。本章概述了一种高效的、基于体外水手转座子的方法，该方法利用这种生物体的自然转化特性来鉴定必需基因。此外，我们还描述了条件表达系统的策略，这些策略将有助于对这类基因进行进一步分析。最后，我们概述了一种基于流感嗜血杆菌中用于鉴定必需基因的方法，该方法可应用于其他不能自然转化的细菌。